Stevia composition

ABSTRACT

Stevia  compositions are prepared from steviol glycosides of  Stevia rebaudiana  Bertoni. The compositions are able to provide a superior taste profile and can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals.

BACKGROUND OF THE INVENTION Field of the Invention

The invention relates to a process for producing a highly purified foodingredient from the extract of the Stevia rebaudiana Bertoni plant andits use in various food products and beverages.

Description of the Related Art

Sugar alternatives are receiving increasing attention due to theawareness of many diseases associated with the consumption of high-sugarfoods and beverages. However, many artificial sweeteners such as dulcin,sodium cyclamate and saccharin have been banned or restricted in somecountries due to concerns about their safety. As a result, non-caloricsweeteners of natural origin are becoming increasingly popular. Thesweet herb Stevia rebaudiana Bertoni produces a number of diterpeneglycosides which feature high intensity sweetness and sensory propertiessuperior to those of many other high potency sweeteners.

The above-mentioned sweet glycosides, have a common aglycon, steviol,and differ by the number and type of carbohydrate residues at the C13and C19 positions. The leaves of Stevia are able to accumulate up to10-20% (on dry weight basis) steviol glycosides. The major glycosidesfound in Stevia leaves are rebaudioside A (2-10%), stevioside (2-10%),and rebaudioside C (1-2%). Other glycosides such as rebaudioside B, D,E, and F, steviolbioside and rubusoside are found at much lower levels(approx. 0-0.2%).

Two major glycosides—stevioside and rebaudioside A (reb A), wereextensively studied and characterized in terms of their suitability ascommercial high intensity sweeteners. Stability studies in carbonatedbeverages confirmed their heat and pH stability (Chang S. S., Cook, J.M. (1983) Stability studies of stevioside and rebaudioside A incarbonated beverages. J Agric. Food Chem. 31: 409-412.)

Steviol glycosides differ from each other not only in their molecularstructures, but also by their taste properties. Usually stevioside isfound to be 110-270 times sweeter than sucrose, rebaudioside A between150 and 320 times sweeter than sucrose, and rebaudioside C between 40-60times sweeter than sucrose. Dulcoside A is 30 times sweeter thansucrose. Rebaudioside A has the least astringent, the least bitter, andthe least persistent aftertaste, thus possessing the most favorablesensory attributes in major steviol glycosides (Tanaka O. (1987)Improvement of taste of natural sweetners. Pure Appl. Chem. 69:675-683;Phillips K. C. (1989) Stevia: steps in developing a new sweetener. In:Grenby T. H. ed. Developments in sweeteners, vol. 3. Elsevier AppliedScience, London. 1-43.) The chemical structure of rebaudioside A isshown in FIG. 1.

Methods for the extraction and purification of sweet glycosides from theStevia rebaudiana plant using water or organic solvents are describedin, for example, U.S. Pat. Nos. 4,361,697; 4,082,858; 4,892,938;5,972,120; 5,962,678; 7,838,044 and 7,862,845.

However, even in a highly purified state, steviol glycosides stillpossess undesirable taste attributes such as bitterness, sweetaftertaste, licorice flavor, etc. One of the main obstacles for thesuccessful commercialization of stevia sweeteners are these undesirabletaste attributes. It was shown that these flavor notes become moreprominent as the concentration of steviol glycosides increases (PrakashI., DuBois G E., Clos J. F., Wilkens K. L., Fosdick L. E. (2008)Development of rebiana, a natural, non-caloric sweetener. Food Chem.Toxicol., 46, S75-S82.).

Rebaudioside B (CAS No: 58543-17-2), or reb B, also known as steviosideA₄ (Kennelly E. J. (2002) Constituents of Stevia rebaudiana In Stevia:The genus Stevia, Kinghorn A. D. (Ed), Taylor & Francis, London, p. 71),is one of the sweet glycosides found in Stevia rebaudiana. Sensoryevaluations show that reb B was approximately 300-350 times sweeter thansucrose, while for reb A this value was approximately 350-450 (Crammer,B. and Ikan, R. (1986) Sweet glycosides from the Stevia plant. Chemistryin Britain 22, 915-916, and 918). The chemical structure of rebaudiosideB is shown in FIG. 2.

It was believed that reb B forms from the partial hydrolysis ofrebaudioside A during the extraction process (Kobayashi, M., Horikawa,S., Degrandi, I. H., Ueno, J. and Mitsuhashi, H. (1977) Dulcosides A andB, new diterpene glycosides from Stevia rebaudiana. Phytochemistry 16,1405-1408). However, further research has shown that reb B occursnaturally in the leaves of Stevia rebaudiana and is currently one ofnine steviol glycosides recognized by FAO/JECFA (United Nations' Foodand Agriculture Organization/Joint Expert Committee on Food Additives)in calculating total steviol glycosides' content in commercial steviolglycoside preparations (FAO JECFA (2010) Steviol Glycosides, Compendiumof Food Additive Specifications, FAO JECFA Monographs 10, 17-21).

Only a few methods are described in literature for preparing reb B.

Kohda et al., (1976) prepared reb B by hydrolysis of reb A withhesperidinase. Reb B was also prepared by alkaline saponification of rebA. The said saponification was conducted in 10% potassiumhydroxide-ethanol. The solution was acidified with acetic acid, andextracted with n-butanol. The butanol layer was washed with water andconcentrated at low temperature in vacuo. The residue was crystallizedfrom methanol to give reb B. (Kohda, H., Kasai, R., Yamasaki, K.,Murakami, K. and Tanaka, O. (1976) New sweet diterpene glucosides fromStevia rebaudiana. Phytochemistry 15, 981-983). The described processesmight be suitable for laboratory scale preparation of reb B, but are notsuitable for any large scale or commercial reb B preparation.

Ahmed et al., used mild alkaline hydrolysis of reb A to prepare reb B.According to the described procedure, reb A was hydrolyzed to reb B byrefluxing with 10% aqueous KOH at 100° C. for 1 hr. After neutralizationwith glacial acetic acid, the precipitated substance was recrystallizedtwice from methanol (Ahmed M. S., Dobberstein R. H., and Farnsworth N.R. (1980) Stevia rebaudiana: I. Use of p-bromophenacyl bromide toenhance ultraviolet detection of water-soluble organic acids(steviolbioside and rebaudioside B) in high-performance liquidchromatographic analysis, J. Chromatogr., 192, 387-393).

The use of methanol as recrystallization media as described in theliterature will require its subsequent removal from the product. It isnoted that handling of toxic substances such as methanol requiresspecialized manufacturing installations and, when applied in foodprocessing, sophisticated food safety measures.

It is also noted that no significant work has been conducted todetermine the potential of reb B as a sweetener or food ingredient.Moreover, reb B is often viewed as process artifact and unnecessaryimpurity in commercial steviol glycosides preparations. No significantevaluation of the influence of reb B on the overall taste profile ofsteviol glycoside preparations has been conducted.

The water solubility of reb B is reported to be about 0.1% (Kinghorn A.D. (2002) Constituents of Stevia rebaudiana In Stevia: The genus Stevia,Kinghorn A. D. (Ed), Taylor & Francis, London, p. 8). In many foodprocesses where highly concentrated ingredients are used, a highlysoluble form of reb B will be necessary.

Considering the facts mentioned above, there is a need to evaluate reb Bas a sweetener and food ingredient and to develop a simple and efficientprocess for food grade reb B preparations suitable for food and otherapplications.

It is further noted that by having a carboxyl group in the moleculemakes it possible for reb B to exist in the forms of various carboxylatesalts. Prior to this invention, it is not believed that differentcarboxylate salt form of reb B had been prepared or evaluated for theirimpact on taste profiles.

Within the description of this invention we will show that, when appliedin specific manner, carboxyate salts of reb B may impact the tasteprofile and offer significant advantages for the use of steviasweeteners in various applications.

SUMMARY OF THE INVENTION

The present invention is aimed to overcome the disadvantages of existingStevia sweeteners. The invention describes a process for producing ahigh purity food ingredient from the extract of the Stevia rebaudianaBertoni plant and use thereof in various food products and beverages asa sweetness and flavor modifier.

The invention, in part, pertains to an ingredient comprising steviolglycosides of Stevia rebaudiana Bertoni plant. The steviol glycodsidesare selected from the group consisting of stevioside, rebaudioside A(FIG. 1), rebaudioside B (FIG. 2), rebaudioside C, rebaudioside D,rebaudioside E, rebaudioside F, dulcoside A, steviolbioside, rubusoside,as well as other steviol glycosides found in Stevia rebaudiana Bertoniplant and mixtures thereof.

The invention, in part, pertains to a process for producing aningredient containing rebaudioside B, and stevioside, rebaudioside A,rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F,dulcoside A, steviolbioside, rubusoside, as well as other steviolglycosides found in Stevia rebaudiana Bertoni plant and mixturesthereof.

In the invention, rebaudioside A commercialized by PureCircle Sdn. Bhd.(Malaysia), containing, rebaudioside A (about 95-100%), stevioside(about 0-1%), rebaudioside C (about 0-1%), rebaudioside F (about 0-1%),rebaudioside B (about 0.1-0.8%), rebaudioside D (about 0-1%), and otherglycosides amounting to total steviol glycosides' content of at least95%, may be used as a starting material. Alternatively stevia extractswith different ratios of steviol glycosides may be used as startingmaterials.

In one embodiment, the starting material is subjected to partial or fullconversion into reb B using an alkaline conversion process. The obtainedreb B is then transformed into a carboxylate salt form by interactingwith the respective base. These salt forms are utilized in similarmanner as “original” reb B with a carboxyl group.

The obtained reb B or its carboxylate salt forms, and mixtures thereofmay be subjected to additional thermal treatment to increase solubility.

The obtained products were applied in various foods and beverages assweeteners, sweetener enhancers and flavor modifiers, including softdrinks, ice cream, cookies, bread, fruit juices, milk products, bakedgoods and confectionary products.

It is to be understood that both the foregoing general description andthe following detailed description are exemplary and explanatory and areintended to provide further explanation of the invention as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings are included to provide a furtherunderstanding of the invention. The drawings illustrate embodiments ofthe invention and together with the description serve to explain theprinciples of the embodiments of the invention.

FIG. 1 shows the chemical structure of rebaudioside A.

FIG. 2 shows the chemical structure of rebaudioside B

FIG. 3 shows an HPLC chromatogram of a stevia composition comprisingrebaudioside A and rebaudioside B.

DETAILED DESCRIPTION OF THE INVENTION

Advantages of the present invention will become more apparent from thedetailed description given hereinafter. However, it should be understoodthat the detailed description and specific examples, while indicatingpreferred embodiments of the invention, are given by way of illustrationonly, since various changes and modifications within the spirit andscope of the invention will become apparent to those skilled in the artfrom this detailed description.

Rebaudioside A commercialized by PureCircle Sdn. Bhd. (Malaysia),containing, rebaudioside A (about 95-100%), stevioside (about 0-1%),rebaudioside C (about 0-1%), rebaudioside D (about 0-1%), rebaudioside F(about 0-1%), rebaudioside B (about 0.1-0.8%) and other glycosidesamounting to total steviol glycosides' content of at least about 95%,may be used as a starting material. Alternatively stevia extracts withdifferent ratios of steviol glycosides may be used as startingmaterials.

The HPLC analysis of the raw materials and products can be performed onan Agilent Technologies 1200 Series (USA) liquid chromatograph, equippedwith Phenomenex Prodigy ODS3, 5 (4.6×250 mm) column at 40° C. The mobilephase was 32:68 mixture of acetonitrile and 10 mmol/L sodium phosphatebuffer (about pH 2.6) at 1 mL/min. A diode array detector set at 210 nmcan be used as the detector. One example of an HPLC chromatogram thusobtained is shown in FIG. 3.

As used herein, unless specified further, “reb B” and “reb Bcomposition” shall be used interchangeably to refer to purifiedrebaudioside B or rebaudioside B in combination with any other chemicalentity.

Preparation of Reb B 1. Alkaline Conversion

One way to obtain reb B starting with reb A is described as follows. RebA is dispersed in aqueous alkaline solution. The concentration of reb Ais about 0-50% (w/v) preferably about 10-25%. The preferred alkalineagents include potassium hydroxide and sodium hydroxide, however otheragents capable of increasing the pH of the media above about pH 7 may beused as well or alternatively. The concentration of alkaline agents isabout 0.05-2.0M, preferably about 0.1-1.0M. The mixture is incubated atabout 10-150° C., preferably about 30-100° C., for a period of about0.5-48 hrs, preferably about 1-24 hrs. As a result, reb A is hydrolyzedto reb B. The molar yield of conversion of reb B is about 5-100%,preferably about 10-90%.

After the reaction, the alkaline agent is neutralized by an acid,preferably by sulfuric acid or ortho-phosphoric acid until a pH of about3.0-5.0 is reached, preferably until a pH of about 3.0-4.0 is reached.Upon neutralization, a precipitate is formed. The precipitate isseparated by any method known in the art such as filtration orcentrifugation and washed with water until the water reaches a pH ofabout 4.0-5.0. The obtained crystalline material is dried under vacuumat about 60-105° C. to yield a mixture of reb A and reb B having a ratioof about 5%:95% to about 95%:5% (w/w), preferably about 50%:50% to about90%:10% (w/w).

2. Optional Post-Conversion Purification

To obtain purified reb B, in one embodiment the separated precipitatedescribed above is suspended in water and the mixture is subjected tocontinuous agitation over about 0.5-24 hrs, preferably about 1-3 hours,at about 50-100° C., preferably about 60-80° C. The ratio of precipitateto water (w/v) is about 1:5 to about 1:20, preferably about 1:10 toabout 1:15. The washed crystals are separated and dried under vacuum atabout 60-105° C. to yield reb B with about 99% purity.

3. Optional Post-Conversion Solubility Enhancement

The following procedure can be used to increase the water solubility ofreb B or any reb B composition. The obtained compositions generally havea water solubility of less than about 0.2% (w/v). In order to increasethe solubility of these compositions, the reb B composition is combinedwith the water at ratio of about 1:1 (w/w) and the obtained mixture issubjected to a gradient heat treatment which results in a high stabilityand high concentration solution. The gradient of about 1° C. per minuteis used in heating the mixture. The mixture is heated to the temperatureof about 110-140° C., preferably about 118-125° C. and is held atmaximum temperature for about 0-120 min, preferably about 50-70 min.After the heat treatment, the solution is cooled down to roomtemperature at gradient of about 1° C. per minute. The solution is spraydried by a laboratory spray drier operating at about 175° C. inlet andabout 100° C. outlet temperatures. An amorphous form of the compositionis obtained with greater than about 20% solubility in water at roomtemperature.

Preparation of Reb B Salt

Reb B obtained using the processes described above, or any otherprocess, can be fully or partially converted into a carboxylate saltform. Reb B, or a composition containing reb B, preferably reb B with apurity greater than 90% (w/w), is dispersed in the water to make anaqueous dispersion with 5-50% (w/v), preferably 5-15% solids content.Excess base is added to achieve a pH level of 6.5-14.0, preferably8.5-11.0. The obtained mixture is incubated for 0.1-24 hrs, preferably1-3 hrs. Then the suspended solids are separated by filtration andwashed with water until neutral pH of the washing water is achieved.Alternatively other reactions able to convert reb B into a carboxylatesalt form may be used. The preferred cations are K⁺ and Na⁺, and therespective bases—KOH and NaOH—are used. However other carboxylate saltsof reb B can be prepared in a similar manner by using the basecorresponding to the desired carboxylate salt.

The obtained reb B carboxylate salt can be further purified and itssolubility enhanced as described above.

While not intending to be bound by theory, it is believed that anysteviol glycoside having a carboxyl group can be converted into itscarboxylate salt form for improving the taste and/or sweetness profileof the molecule. Another example of a steviol glycoside containing acarboxyl group is steviolbioside.

Use of Reb B Carboxylate Salt

The reb B carboxylate salt described above can be used as a sweetnessenhancer, a flavor enhancer and/or a sweetener in various food andbeverage products. Non-limiting examples of food and beverage productsinclude carbonated soft drinks, ready to drink beverages, energy drinks,isotonic drinks, low-calorie drinks, zero-calorie drinks, sports drinks,teas, fruit and vegetable juices, juice drinks, dairy drinks, yoghurtdrinks, alcohol beverages, powdered beverages, bakery products, cookies,biscuits, baking mixes, cereals, confectioneries, candies, toffees,chewing gum, dairy products, flavored milk, yoghurts, flavored yoghurts,cultured milk, soy sauce and other soy base products, salad dressings,mayonnaise, vinegar, frozen-desserts, meat products, fish-meat products,bottled and canned foods, tabletop sweeteners, fruits and vegetables.

Additionally the reb B carboxylate salt compositions can be used in drugor pharmaceutical preparations and cosmetics, including but not limitedto toothpaste, mouthwash, cough syrup, chewable tablets, lozenges,vitamin preparations, and the like.

The compositions can be used “as-is” or in combination with othersweeteners, flavors and food ingredients.

Non-limiting examples of sweeteners include steviol glycosides,stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudiosideD, rebaudioside E, rebaudioside F, dulcoside A, steviolbioside,rubusoside, as well as other steviol glycosides found in Steviarebaudiana Bertoni plant and mixtures thereof, stevia extract, Luo HanGuo extract, mogrosides, high-fructose corn syrup, corn syrup, invertsugar, fructooligosaccharides, inulin, inulooligosaccharides, couplingsugar, maltooligosaccharides, maltodextins, corn syrup solids, glucose,maltose, sucrose, lactose, aspartame, saccharin, sucralose, sugaralcohols.

Non-limiting examples of flavors include lemon, orange, fruit, banana,grape, pear, pineapple, bitter almond, cola, cinnamon, sugar, cottoncandy, vanilla flavors.

Non-limiting examples of other food ingredients include flavors,acidulants, organic and amino acids, coloring agents, bulking agents,modified starches, gums, texturizers, preservatives, antioxidants,emulsifiers, stabilisers, thickeners, gelling agents.

The following examples illustrate various embodiments of the invention.It will be understood that the invention is not limited to thematerials, proportions, conditions and procedures set forth in theexamples, which are only illustrative.

Example 1 Preparation of Stevia Composition

100 g of rebaudioside A produced by PureCircle Sdn. Bhd. (Malaysia),containing, 98.1% rebaudioside A, 0.3% stevioside, 0.2 rebaudioside C,0.2% rebaudioside F, 0.4% rebaudioside B and 0.6% rebaudioside D wasdispersed in 1000 mL aqueous KOH (1M) and incubated at 50° C. for 2hours. The mixture temperature was decreased to 20° C. and the pH wasadjusted to pH 4.0 with sulfuric acid. The solution was held undermoderate agitation conditions for 4 hours and a precipitate was formed.The precipitate was filtered and washed on the filter with 2000 mL ofwater. The washed crystals were dried under vacuum to yield 86 gmaterial containing about 84% reb A and 16% reb B. The water solubility(at 25° C.) of obtained material was about 0.2% (w/v).

Example 2 Preparation of Stevia Composition

100 g of rebaudioside A produced by PureCircle Sdn. Bhd. (Malaysia),containing, 98.1% rebaudioside A, 0.3% stevioside, 0.2 rebaudioside C,0.2% rebaudioside F, 0.4% rebaudioside B and 0.6% rebaudioside D wasdispersed in 1000 mL aqueous KOH (1M) and incubated at 80° C. for 5hours. The mixture temperature was decreased to 20° C. and the pH wasadjusted to about pH 4.0 with sulfuric acid. The solution was held undermoderate agitation conditions for 4 hours and a precipitate was formed.The precipitate was filtered and washed on the filter with 2000 mL ofwater. The washed crystals were dried under vacuum to yield about 75 gmaterial containing about 9% reb A and about 91% reb B. The watersolubility (at 25° C.) of obtained material was about 0.1% (w/v).

Example 3 Preparation of Stevia Composition

100 g of rebaudioside A produced by PureCircle Sdn. Bhd. (Malaysia),containing, 98.1% rebaudioside A, 0.3% stevioside, 0.2 rebaudioside C,0.2% rebaudioside F, 0.4% rebaudioside B and 0.6% rebaudioside D wasdispersed in 1000 mL aqueous KOH (1M) and incubated at 80° C. for 7hours. The mixture temperature was decreased to 20° C. and the pH wasadjusted to about pH 4.0 with sulfuric acid. The solution was held undermoderate agitation conditions for 3-4 hours and a precipitate wasformed. The precipitate was filtered and washed on the filter with 2000mL of water. The washed crystals were dried under vacuum to yield about71 g material containing about 99.1% reb B. The water solubility (at 25°C.) of obtained material was about 0.1% (w/v).

Example 4 Preparation of Reb B

75 g of material prepared according to EXAMPLE 2 was suspended in 1000mL water. The mixture temperature was increased to 70° C. The suspensionwas held under moderate agitation conditions for 4 hours. The crystalswere filtered and dried under vacuum to yield about 65 g materialcontaining about 99.0% reb B. The water solubility (at 25° C.) ofobtained material was about 0.1% (w/v).

Example 5 Preparation of Reb B Carboxylate Salt

100 g of reb B prepared according to EXAMPLE 4 was suspended in 1000 mLwater. The suspension was held under moderate agitation conditions atroom temperature for 1 hr. The pH of the mixture was adjusted to pH 11.0with 1M KOH solution. The obtained mixture was agitated for 3 hrs. Thecrystals were filtered and washed with water on the filter until thewashing water reached a neutral pH. The obtained crystalline materialwas dried under vacuum to yield about 95 g material. The watersolubility (at 25° C.) of obtained material was about 0.1% (w/v).

Example 6 Preparation of Soluble Stevia Composition

50 g material prepared according to EXAMPLE 1 was mixed with 50 g ofwater and incubated in thermostatted oil bath. The temperature wasincreased at 1° C. per minute to 121° C. The mixture was maintained at121° C. for 1 hour and then the temperature was decreased to roomtemperature (25° C.) at 1° C. per minute. The solution was dried usingYC-015 laboratory spray drier (Shanghai Pilotech Instrument & EquipmentCo. Ltd., China) operating at 175° C. inlet and 100° C. outlettemperature. About 47 g of an amorphous powder was obtained with about25% (w/v) solubility in water (at 25° C.).

Example 7 Preparation of Soluble Stevia Composition

42 g of reb A produced by PureCircle Sdn. Bhd. (Malaysia) with purity of99.2% (dry basis) and 8 g of reb B prepared according to EXAMPLE 4 weremixed with 50 g of water and incubated in thermostatted oil bath. Thetemperature was increased at 1° C. per minute to 121° C. The mixture wasmaintained at 121° C. for 1 hour and then the temperature was decreasedto room temperature (25° C.) at 1° C. per minute. The solution was driedusing YC-015 laboratory spray drier (Shanghai Pilotech Instrument &Equipment Co. Ltd., China) operating at 175° C. inlet and 100° C. outlettemperature. About 48 g of an amorphous powder was obtained with about1.5% (w/v) solubility in water (at 25° C.).

Example 8 Preparation of Soluble Stevia Composition

42 g of reb A produced by PureCircle Sdn. Bhd. (Malaysia) with purity of99.2% (dry basis) and 8 g of reb B potassium salt prepared according toEXAMPLE 5 were mixed with 50 g of water and incubated in thermostattedoil bath. The temperature was increased at 1° C. per minute to 121° C.The mixture was maintained at 121° C. for 1 hour and then thetemperature was decreased to room temperature (25° C.) at 1° C. perminute. The solution was dried using YC-015 laboratory spray drier(Shanghai Pilotech Instrument & Equipment Co. Ltd., China) operating at175° C. inlet and 100° C. outlet temperature. About 49 g of an amorphouspowder was obtained with about 2.5% (w/v) solubility in water (at 25°C.).

Example 9 Low-Calorie Orange Juice Drink

Orange concentrate (35%), citric acid (0.35%), ascorbic acid (0.05%),orange red color (0.01%), orange flavor (0.20%), and 0.05% steviacomposition, were blended and dissolved completely in water (up to 100%)and pasteurized. The stevia composition was selected from a commercialstevia extract (containing stevioside 26%, rebaudioside A 55%, and 16%of other glycosides), a commercial rebaudioside A (containing 98.2% rebA) or material obtained according to EXAMPLE 7 and EXAMPLE 8.

The sensory evaluations of the samples are summarized in Table 1. Thedata show that the best results can be obtained by using the compositionobtained according to EXAMPLE 8. Particularly the drinks prepared withsaid composition exhibited a rounded and complete flavor profile andmouthfeel.

TABLE 1 Evaluation of orange juice drink samples Comments Sample FlavorAftertaste Mouthfeel Stevia Extract Sweet, licorice notes Bitterness andNot acceptable aftertaste Reb A Sweet, slight licorice notes Slightbitterness and Not acceptable aftertaste EXAMPLE 7 High qualitysweetness, Clean, almost no Full pleasant taste almost similar tobitterness and slight sucrose, fairy rounded and aftertaste balancedflavor EXAMPLE 8 High quality sweetness, Clean, no bitterness Fullpleasant taste similar to sucrose, and no aftertaste rounded andbalanced flavor

The same method can be used to prepare juices and juice drinks fromother fruits, such as apples, lemons, apricots, cherries, pineapples,mangoes, etc.

Example 10 Zero-Calorie Carbonated Beverage

Carbonated beverages made according to the formulas presented in Table 2were prepared.

TABLE 2 Carbonated Beverage Formulas Quantity, % Stevia IngredientsExtract Reb A EXAMPLE 7 EXAMPLE 8 Cola flavor 0.340 0.340 0.340 0.340ortho-Phosphoric 0.100 0.100 0.100 0.100 acid Sodium citrate 0.310 0.3100.310 0.310 Sodium benzoate 0.018 0.018 0.018 0.018 Citric acid 0.0180.018 0.018 0.018 Stevia composition 0.050 0.050 0.050 0.050 Carbonatedwater to 100 to 100 to 100 to 100

The sensory properties were evaluated by 20 panelists. The results aresummarized in Table 3.

TABLE 3 Evaluation of zero-calorie carbonated beverage samples Number ofpanelists detected the attribute Taste attribute Stevia Extract Reb AEXAMPLE 7 EXAMPLE 8 Bitter taste 15 10 2 0 Astringent taste 16  9 1 0Aftertaste 14 12 2 0 Comments Quality of sweet taste Bitter aftertaste(15 Bitter aftertaste (10 Clean (17 of 20) Clean (20 of 20) of 20) of20) Overall evaluation Satisfactory (1 of Satisfactory (5 ofSatisfactory (16 of Satisfactory (20 of 20) 20) 20) 20)

The above results show that the beverages prepared using the compositionobtained according to EXAMPLE 8 possessed the best organolepticcharacteristics.

Example 11 Diet Cookies

Flour (50.0%), margarine (30.0%) fructose (10.0%), maltitol (8.0%),whole milk (1.0%), salt (0.2%), baking powder (0.15%), vanillin (0.1%)and different stevia compositions (0.03%) were kneaded well indough-mixing machine. The obtained dough was molded and baked in oven at200° C. for 15 minutes. The stevia compositions were selected from acommercial stevia extract (containing stevioside 26%, rebaudioside A55%, and 16% of other glycosides), a commercial rebaudioside A(containing 98.2% reb A) and material obtained according to obtainedaccording to EXAMPLE 7 and EXAMPLE 8.

The sensory properties were evaluated by 20 panelists. The best resultswere obtained in samples containing the composition obtained accordingto EXAMPLE 8. The panelists noted a rounded and complete flavor profileand mouthfeel.

Example 12 Yoghurt

Different stevia compositions (0.03%) and sucrose (4%) were dissolved inlow fat milk. The stevia compositions were selected from a commercialstevia extract (containing stevioside 26%, rebaudioside A 55%, and 16%of other glycosides), a commercial rebaudioside A (containing 98.2% rebA) and the material obtained according to EXAMPLE 7 and EXAMPLE 8. Afterpasteurizing at 82° C. for 20 minutes, the milk was cooled to 37° C. Astarter culture (3%) was added and the mixture was incubated at 37° C.for 6 hours then at 5° C. for 12 hours.

The sensory properties were evaluated by 20 panelists. The best resultswere obtained in samples containing the composition obtained accordingto EXAMPLE 8. The panelists noted a rounded and complete flavor profileand mouthfeel.

It is to be understood that the foregoing descriptions and specificembodiments shown herein are merely illustrative of the best mode of theinvention and the principles thereof, and that modifications andadditions may be easily made by those skilled in the art withoutdeparting for the spirit and scope of the invention, which is thereforeunderstood to be limited only by the scope of the appended claims.

1. A stevia composition comprising a carboxylate salt of rebaudioside B,made by a process comprising the steps of: providing a stevia sweetener;providing an alkaline aqueous solution; dispersing the stevia sweetenerin the alkaline solution and incubating for about 12 to 48 hours atabout 55-75° C. to form a mixture; cooling the mixture to about 10-30°C. and adjusting the pH with acid to about pH 3.0-4.0; incubating themixture to obtain a precipitate; separating the precipitate and washingit with water; drying the washed precipitate to obtain the steviacomposition; dispersing the stevia composition in water to form amixture; adding a base corresponding to the carboxylate salt into themixture to achieve a pH level of 6.5 to 14.0; incubating the mixture for0.1 to 24 hours to facilitate conversion of the rebaudioside B into thecarboxylate salt of rebaudioside B; and separating and drying the steviacomposition comprising the carboxylate salt of rebaudioside B. 2-5.(canceled)
 6. The stevia composition of claim 1, the compositioncomprising a mixture of rebaudioside A, rebaudioside B, and rebaudiosideB caboxylate salt, wherein the ratio of rebaudioside A to rebaudioside Band rebaudioside B carboxylate salt ranges from about 5%:95% to about95%:5% (w/w).
 7. The stevia composition of claim 1, the compositioncomprising a mixture of rebaudioside D, rebaudioside B, and rebaudiosideB carboxylate salt, wherein the ratio of rebaudioside D to rebaudiosideB and rebaudioside B carboxylate salt ranges from about 5%:95% to about95%:5% (w/w). 8-11. (canceled)
 12. A food, beverage, pharmaceutical orcosmetic composition comprising the stevia composition of claim 1.